Tumors produce circulating factors that have the potential to be used as diagnostic markers for the presence of the tumor. These markers may be tumor type specific, angiogenic factors, or cytokines involved in the immune response to the tumor. The diagnostic based on just one marker yield too many false positives. What is needed is the ability to assay many markers (up to 100 markers) by proteomics and then develop an accurate correlation between the presence of the markers and the presence of the tumor, thus eliminating false positives. We propose use of the supersensitive MultiPhoton Detection (MPD) enhanced to immunoassays (IA/MPD) and protein-chips (P-chips/MPD), to detect tumor markers in concentration ranges from 0.01 pg/ml level to 500 pg/ml. The technology will result in a more sensitive and specific diagnostic method, which may enable the detection of the tumors in an earlier stage of development. The objective of Phase I of this proposal is to develop the supersensitive IA/MPD assay techniques for the angiogenesis factors, Fibroblast Growth Factor-1 (FGF-1) that have been found to be expressed in many tumor types. Then clinically relevant serum samples will be screened for FGF-1, along with FGF-2, Vascular Endothelial Growth Factor (VEGF), Hepatocyte Growth Factor (HGF), IL-lbeta, IL-6, IL-10, IL-11, IL-12, and TNF-alpha, and tumor markers like Prostate Serum Antigen. Ultrasensitive IA/MPD assays currently exist for many of these factors. The clinically relevant serum samples will include pre- and post- cancer treatment patients, and healthy control subjects. Post treatment cancer patients will be monitored over time to see if marker level variations over time can predict tumor recurrence. Results will be examined to determine if a particular pattern of marker levels correlates with the presence of a tumor. The goal of Phase II is the development of the MPD enhanced P-chip. The P-chips/MPD will be adapted to a large set of markers including PSA, CEA, CA-125, and alpha-fetoprotein, cytokines and angiogenesis factors that correlate with the presence of a tumor. Additional factors, which are found expressed by tumors, but can not be detected by current relatively low sensitivity diagnostics will also be studied. Testing on clinically relevant samples will be expanded. Once the importance of concurrent measurement of several of these biomarkers are confirmed, we will develop dedicated supersensitive P-chips for quantifying up to 100 targeted marker proteins with detection ranges from 0.01 to 500 pg/ml. Thus, low cost cancer diagnostics and therapy monitoring assay will become available and a potential mechanism(s) may be suggested for earlier stage tumor formation since some of the markers may only be expressed at lower levels at the early stages.